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Novel aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, of the ambrosia

beetle, Platypus quercivorus (Coleoptera: Platypodidae)

Article · January 2007

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「森林総合研究所研究報告」(Bulletin of FFPRI), Vol.6, No.1 (No.402), 49 - 57, March, 2007

論 文（Original article）

Novel aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, of the

ambrosia beetle, Platypus quercivorus (Coleoptera: Platypodidae)

TOKORO Masahiko 1)*, KOBAYASHI Masahide 2), SAITO Shoichi 3),

KINUURA Haruo 4), NAKASHIMA Tadakazu1), SHODA – KAGAYA Etsuko 1),
KASHIWAGI Takehiro5), TEBAYASHI Shin-ichi5), KIM Chul-Sa5)
and MORI Kenji 6)
Abstract
We identified an aggregation pheromone of Platypus quercivorus using volatiles from the boring frass of an
unmated male. Gas chromatography – electroantennographic detector (GC–EAD) showed an EAD active component
from the volatiles. GC – mass spectrometry (MS) data and comparison of the retention times of stereospecific
synthetic compounds of the optical isomers on chiral GC column showed that the chemical structure of the EAD
active component was (1S,4R)-p-menth-2-en-1-ol (quercivorol). A field trapping tests using quercivorol and its
optical isomer demonstrated that the quercivorol is main component of aggregation pheromone of P. quercivorus.

Key words : Platypus quercivorus, aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, quercivorol, ambrosia beetle

Introduction
Platypus quercivorus (Murayama)(Coleoptera: Platypodidae)
is an ambrosia beetle that bore into the trunks of oaks and other
broad-leaved trees in Japan, Taiwan, India, Indonesia and New
Guinea (Wood & Bright, 1992). In platypodid ambrosia beetles,
male-initiated monogamy is usual (Kirkendall, 1983). Mass
mortality of oaks, especially Quercus crispula Blume, resulting
when large infestations of these beetles spread Japanese oak
wilt, has occurred continuously in Japan since 1934 (Hijii et
al.,1991; Ito & Yamada, 1998). Ito et al., (1998) reported that
ambrosia fungi caused the mass mortality of Japanese oak, and
that P. quercivorus is considered to be a critical vector of the
fungi. Later, a phytopathogenic fungus was isolated from the
gallery of the trunk and mycangia of the beetle, and named as
Raffaelea quercivora (Kubono & Ito, 2002).
In recent studies, concentrative boring was observed on a
particular oak tree soon after the initial attack of the beetle (Ueda
& Kobayashi, 2001, Kobayashi & Ueda, 2003), suggesting
the presence of semiochemicals attracting a large number of
individuals. Field experiments have indicated that chemical
communication was responsible for mass attacks on oaks by the
beetles at long range (Ueda & Kobayashi, 2005), whereas sound
communication is a cue for their mating behavior at close range
as well (Ohya & Kinuura, 2001, Kobayashi & Ueda, 2002).
In some platypodid species that attack their hosts in large
numbers, the males release aggregation pheromone, which
attracts conspecific males and females (Madrid et al., 1972;
Milligan, 1982; Milligan & Ytsma, 1988; Milligan et al., 1988).
Sulcatol (6-methyl-5-hepten-2-ol), which is the aggregation
pheromone of some scolytid beetles including Gnathotricus
sulcatus (LeConte) (Byrne et al.,1974), G. sulcatus materiarius
Fitch (Fletchmann & Berisford, 2003) and G. sulcatus retusus
(LeConte) (Borden et al.,1979), was determined to be the main
component of aggregation pheromones of platypodid beetles
as P. flavicornis (F.) (Renwick et al., 1977) and P. wilsoni
(Shore & McLean, 1983). Although Ueda and Kobayashi
(2001) suggested the presence of aggregation pheromone
in P. quercivorus, it has not been chemically identified and
biologically assayed. In this paper, we identify the aggregation
pheromone of P. quercivorus.
Materials and Methods
Test Insects
Logs of Q. crispula Blume infested by P. quersivorus were
collected from oak forests in Aizu-wakamatsu, Fukushima
prefecture and Keihoku, Kyoto city, Kyoto prefecture, Japan in

原稿受付：平成 18 年 12 月 11 日 Received Dec. 11, 2006  原稿受理：平成 19 年 1 月 9 日 Accepted Jan. 9, 2007

* Department of Forest Entomology, Forestry and Forest Products Research Institute (FFPRI), 1 Matsunosato, Tsukuba, Ibaraki 305-8687,
Japan; e-mail: tokoro@affrc.go.jp
1) Department of Forest Entomology, Forestry and Forest Products Research Institute (FFPRI)
2) Kyoto Prefectural Forestry Experimental Station
3) Yamagata Prefectural Forest Research and Instruction Center
4) Kansai Research Center, Forestry and Forest Products Research Institute (FFPRI)
5) Department of Bioresources Science, Faculty of Agriculture, Kochi University
6) Toyo Gosei Co., Ltd., Photosensitive Materials Research Center
50 TOKORO M.et al.

October 2003 and 2004, and kept in a temperature-controlled
room (16°C). Emerging beetles were collected daily, sexed, and
kept until use of experiments.
Chemicals
Synthetic racemic cis-p-menth-2-en-1-ol for 2004 and
2005 field traps test was purchased from Sankei Chemical Co
Ltd. (Tokyo, Japan) as a mixture of cis-p-menth-2-en-1-ol (>
76.2%) and trans-isomer (> 17.9%) (racemic quercivorol I). In
this paper, the cis-isomer is defined as the stereoisomer with
both the methyl and the isopropyl groups in the same direction.
Racemic cis-p-menth-2-en-1-ol, synthesized from (±)-cryptone,
chemical purity was 79.8%, contains 4.1% of trans-p-menth-
2-en-1-ol for 2006 field trap test (racemic quercivorol II: Mori,
2006). (1S,4R)-p-menth-2-en-1-ol (quercivorol: chemical purity
92.0%, optical purity 96.0%) was synthesized from (S)-perillyl
alcohol (Mori, 2006). (1R,4S)-p-menth-2-en-1-ol (chemical
purity 95.4%, optical purity 96.1%) was synthesized from (R)-
limonene (Kashiwagi et al., 2006). Ethyl alcohol was purchased
from Nihon Alcohol Hanbai Corp. Acetone (99.8%) and
n-hexane (96.0%) and all other reagents were purchased from
Wako Chemical Ltd. and Nacalai Tesque Ltd. in Japan.
Collection of volatiles
Logs of Q. serrata Thunb. ex Murray uninfested by P.
quercivorus (20–40 cm long, 10–20 cm i.d.) were soaked in water
for over ten days to improve the breeding condition of the beetles
(Kitajima & Goto, 2004).
Holes (2 mm i.d. and 1–1.5 cm depth) were bored by drill
into the logs at 2–5 cm intervals. Unmated male beetles were
introduced as pioneers into the holes through a plastic pipette
chip (1 ml volume), with tips cut to allow the beetle to pass
into the logs. Each hole occupied by one male beetle. Within
several hours, the male beetles started to push out the frass to
the surrounding tunnel entrances. And after a while, boring
the males deposited droplets from their anuses onto the boring
frass circles. Similar behaviors were reported in males of P.
apicalis and P. gracilis (Milligan & Ytsma, 1988). After four
days, the masses of boring frass from single tunnel entrance
were collected at 10:00 a.m. and put into glass screw vials (1.5–
5 ml). Volatiles from the head space of the vials were collected
using a solid phase microextraction (SPME) holder with a fiber
of 100 μm polydimethylsiloxane (Supelco, Bellefonte, PA,
USA) for about six hours. To extract more volatile components,
boring frass from 30 tunnel entrances of unmated male beetles
were soaked in 3 ml acetone for over one day. An unmated male
or female beetle newly emerged was also soaked in 150 μl of
hexane for over one day for extraction.
Analysis of volatile components
Gas chromatographic–electroantennographic detection (GC–EAD)
Volatiles collected from the acetone boring frass extracts
or SPME fiber were analyzed by GC–EAD system described
by Chen et al., 2006. A Hewlett-Packard 5890 series II GC
was equipped with a nonpolar column HP1-MS (30 m × 0.25
mm i.d. × 0.25 μm film thickness, J&W Scientific, Folsom,
CA, USA), a polar column DB-23 (30 m × 0.25 mm i.d. ×
0.25 μm film thickness, J&W Scientific), or a chiral separation

A Antenna B

Indifferent Electrode
Glass Capillary
0.3mm
Plastic Tip

Glass Capillary
Glue

Recording Electrode
Rubber Septum

Acrylic Holder
�mm

Fig. 1. A: Acrylic antenna holder with electrodes for EAD preparation

  B: Close-up view of antenna. See text for details.

森林総合研究所研究報告 第 6 巻 1 号 , 2007
 Novel aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, of the ambrosia beetle, Platypus quercivorus (Coleoptera: Platypodidae)
column CP-Chirasil-Dex CB (25 m × 0.25 mm i.d. × 0.25 μm
film thickness, Chrompack, London, UK). The temperatures
of the injection port and flame ionization detector (FID) port
were 220°C and 250°C, respectively. Oven temperatures were
programmed at 50°C for 1 min, then 10°C/min to 250°C for the
non-polar columun, 60°C for 1 min, then 15°C / min to 200°C
for the polar column, 60°C for 1 min, then 10°C / min to 200°
C for the chiral column. For SPME sample injection, a 0.75 mm
diameter glass inlet liner (Supelco) was used, and for acetone
extracts sample injection, a 3 mm diameter glass inlet liner was
used for all column types.
An antenna including basic segment was gently pulled
out from a live beetle with tweezers under a stereoscopic
microscope, and was placed between recording and indifferent
electrodes in an acrylic holder (Fig. 1). The acrylic holder that
we used was modified from the method of Nojima et al., (2003)
for tiny antennae from small beetles, using a 1.5 mm diameter
glass capillaries (tip 0.1–0.2 mm i.d.) filled with Grace’s insect
cell culture medium (Invitrogen Corp., Carlsbad, CA, USA)
for electric conductivity. Silver wires were inserted into the
electrodes through acrylic holder at one end and were connected
to a Syntech (Hilversum, the Netherlands) EAG probe through
an interchangeable insert at the other. The identities of EAD
active compounds were verified by matching retention times with
purified synthetic standards injected onto these three columns. An
anntenal EAD response to the identified compound was verified
with 10 ng samples of purified synthetic standards dissolved with
n-hexane, and averaged EAD activity was calculated from five
individual responses to the standards.
GC – mass spectrometry (GC–MS)
The EAD active components were analyzed by GC–MS
(HP6890 GC and HP 5973N MSD, Agilent Technology, Huston,
USA) in EI mode at 70 eV. Ion source temperature of 230°C, Q
pole temperature of 150°C and interface temperature of 250°C.
Samples were analyzed on a non-polar column (HP1-MS), a
polar column (DB23) or a chiral separation column (CP-Chirasil-
DEX CB), under the same conditions as those of GC–EAD
analyses, except for initial temperature (35ºC) and initial holding
time (3 min) in the case of the non-polar column. The GC–EAD
active components were identified by mass spectral matches
to library spectra (Wiley7n: Agilent Technology) followed by
comparison of retention times with those of authentic compounds.
Field trap tests
Field trap tests in 2004 were performed using traps with
cross barriers (30 cm width × 20 cm in height) and water basins
(20 cm i.d.×14 cm in depth) (Sankei Chemical Co. Ltd.,Tokyo,
Japan). Lures for the traps were made from plastic cups (5.8
cm i.d. × 2 cm in depth) containing cotton wicks infused with
Bulletin of FFPRI, Vol.6, No.1, 2007
51
1 ml of synthetic racemic quercivorol Ⅰ (quercivorol), 50 ml
of 70% ethyl alcohol solution (ethanol) or 50 ml of distilled
water (control). Three kinds of traps were baited with racemic
quercivorol Ⅰ , ethanol and control and were set at a height of
1.5 m on trees in forests damaged by P. quercivorus at two sites
in Kushibiki, Yamagata prefecture and at two sites in Keihoku,
Kyoto prefecture, Japan in the summer, 2004. The captured
beetles at each trap were collected every 4 days, during 28
days of tested period and counted for each sex. Lures were
replenished and traps were shifted in each site every 8 days.
In summer of 2005, we tested the effect of dose of synthetic
racemic quercivorol, useing same type of traps and lures with
20, 60, 200 or 600 μl of racemic quercivorol Ⅰ in damaged
forest in Atsumi, Yamagata prefecture (2005). In summer of
2006 we also tested attractance of optical isomers of quecivorol
using funnel traps (Pherotech Inc., Delta, BC, Canada) and
with 20 μl of purified enantiomers ((1S,4R)–isomer or (1R,4S)–
isomer) or racemic quercivorol Ⅱ in damaged forest in Oguni,
Yamagata prefecture (2006). The captured beetles at each trap
were collected every week and counted for each sex.
Statistical analysis for field trap test.
Statistical analyses were performed with SYSTAT ver.
9.01 (SPSS Inc., 1998). In all analyses a Type I error (α) rate of
0.05 was used. First, field data of numbers of captured beetles
at each trap of Atsumi and Keihoku in 2004 were analyzed in
three-way ANOVAs with the lure treatment, test site and period
of the experiment as fixed factors. Because numbers of the
beetles were heteroscedastic, log10 (N+1)–transformations were
performed prior to ANOVA analysis. There were few captured
beetles in one site in Keihoku, therefore we also performed the
same statistical analysis with this site excluded from the dataset.
In cases where no significant interactions between factors
occurred in ANOVA, pairwise differences between factor
combinations were tested with the Bonferroni adjusting method
(SPSS Inc., 1998). We also assessed effective dose of racemic
quercivorol Ⅰ in the data collected at Atsumi in 2005. Because
there were no significant interactions between dose of racemic
quercivorol and collection period, we performed ANCOVAs
without such interactions. Last, we examined repeated measures
ANOVAs for the experiment dealing with the optical isomers of
quercivorol for the field test at Oguni in 2006.
To evaluate sex ratios for the captured beetles in response
to racemic quercivorol Ⅰ and ethanol, we conducted Fisher’s
exact probability tests for two-way tables.
Results and Discussion
Analysis of Volatile component
GC–EAD
Results of GC–EAD analyses are shown in Fig. 2. With
52 TOKORO M.et al.

regards to boring frass volatiles from acetone extracts produced (a)

�� ���������������
by unmated males, the antennae of both sexes responded to a
peak A eluting at retention time 4.55 min and retention index
(RI: 1123) on the non-polar column, 6.82 min (RI: 1577) on ���� ��

the polar column, and ca.10.0 min (RI: 1320) on the chiral ����
(b)
column. EAD response to peak A was 0.29 ± 0.16 mV (mean A
�� �������������
Male antenna

± SD) for female antennae of Fukushima, 0.26 ± 0.19 mV for

female antennae of Kyoto, 0.34 ± 0.32 mV for male antennae FID
B
��
����
of Fukushima, and 0.43 ± 0.25 mV for male antennae of Kyoto. EAD
����
Neither were there no significant differences (t-test, P<0.05) ���������������������
of EAD responses to peak A between the two geographical �
�� ��
locations, nor there were between males and females. However,
female antennal responses were more reproducible than �� � � �
male antennal responses. EAD responses of peak A in boring
frass volatiles from SPME samples were also observed (data ���������������

not shown). It is possible that female antennae have more

� �� � �������������
sensitive sensillae to this component than male antennae. The
results suggest that this component plays important roles as an
aggregation pheromone, for male and female orientation, but ���������������������
there is a possibility that other undetected compounds may play
Fig. 2. Simultaneous recordings of the GC-FID (upper trace)
role as kiromone for male orientation.
and GC–EAD (lower trace) responses to the boring
The mean amount of the peak A was 106.04 ± 30.72ng frass acetone extracts produced by unmated males of P.
from male whole body hexane extracts from Kyoto. quersivorus using a non-polar column.
(a) Female antenna from Kyoto 
Interestingly, we also detected a minute amount (less than 1 ng)
(b) Male antenna from Kyoto
of peak A from female whole body hexane extracts from Aizu- (c) Magnification of the chromatograms of peaks A and B
wakamatu. Hereafter, it will be necessary to clarify whether the A : EAD-active component B: stereo isomer of peak A
female also secretes this component actively.

� A

B
Retention
min.

� A

B
Retention
min.

100
�
Relative intensity

50

Fig. 3. Results of GC–MS analyses of boring frass volatiles extracted by acetone and SPME, using nonpolar column.
(a) Total ion chromatogram of the boring frass volatiles from acetone extract.
(b) Total ion chromatogram of SPME sample.
(c) Mass spectrum of peak A from boring frass acetone extract.
A: EAD-active component  B: trans-stereoisomer of peak A compound
Note that retention times for peaks A (9.27 min) and B (8.95 min) in (a) and (b) were identical,
respectively.

森林総合研究所研究報告 第 6 巻 1 号 , 2007
 Novel aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, of the ambrosia beetle, Platypus quercivorus (Coleoptera: Platypodidae) 53

GC–MS 2005, the number of captured beetles significantly changed

Results of GC–MS analyses of the EAD–active component with dose of racemic quercivorol I (Fig. 6, ANCOVAs, dose
(peak A: Fig. 2a, b) are shown in Fig. 3. of racemic quercivorol I: F 1,7=5.643, P=0.049, test period:
The EI mass spectrum of the peak A compound showed F 1,7=8.228, P=0.024). We suggest that the optimal dose of
a molecular ion at m/z 154 (M+·) and characteristic fragment racemic quercivorol I is above 600 μl / trap, though we cannot
ion peaks at m/z 43(C3H7 : base peak), 55, 69, 81, 93 (M–43 define exact value.
–H2O), 111 (M–43), 121 (M–CH3–H2O), 136 (M–H2O), and From the results of the field trap test at Oguni, Yamagata
139 (M–CH3) (Fig. 3c). The mass spectral matching to library in 2006 using optical isomers, traps with both enantiomers and
spectra of authentic compounds and comparable retention racemic quercivorol II caught the beetles, but in the control
times with among peak A and authentic compounds indicated traps did not (Fig. 7). In total, we captured about twice as
that the active peak was monoterpene alcohol of molecular many beetles with (1S,4R) traps as with (1R,4S) and racemic
weight 154 with a cyclohexene ring, and that the ring had a quercivorol II. However, we cannot detected significant
hydroxyl function on the 1st carbon and an isopropyl function differences between lures (Repeated measured ANOVAs,
on the 4th carbon. We determined the chemical structure of type of lure: F3,16=1.310, P=0.306, test period: F1,16=5.904,
the EAD-active compound to be cis-p-menth-2-en-1-ol. P=0.027, type of lure × test period: F3,16=0.755, P=0.535). The
+·
Peak B also showed a molecular ion at m/z 154 (M ) and captured number of beetles significantly changed between the
similar characteristic fragment ion peaks with peak A, so we test periods. The decreased number of beetles was likely due to
determined the compound to be a trans-p-menth-2-en-1-ol by weather changes between the test periods because the beetles’
comparison of authentic compounds for retention times and dispersal behavior is known to change in response to sunlight
mass spectra. However, no reproducible response to peak B was and wind (Ueda & Kobayashi, 2000).
detected by GC–EAD analyses.
Teble 1. Results of a three-way ANOVA of the effects of lure
Chiral analysis with the authentic compounds by
(quercivorol*, ethanol and water), test period and test site
Kashiwagi et al. (2006) indicates peak A to be (1S,4R)-p-menth- on log-transformed number of beetles collected per trap
2-en-1-ol (Fig. 4), for which we proposed the name quercivorol (a) for all sites , (b) for well-collected sites
after P. quercivorus.     *racemic quercivorol I
(a)
Field trap tests factor df MS F-ratio P
lure 2 1.789 35.876 < 0.0001
Results of field trap test at Yamagata and Kyoto in
test site 3 0.439 8.799 0.001
2004 were shown in Fig. 5. Number of captured beetles by test period 3 0.149 2.986 0.059
quercivorol I and ethanol did not significantly differ (Fisher’s lure × test site 6 0.132 2.649 0.051
exact probability test) among female and males, the numbers of test site × test period 9 0.061 1.215 0.345
both sexes were combined further statistical analyses. Because test period × lure 6 0.062 1.240 0.332
18 0.050
quercivorol traps attracted both sexes, quercivorol apparently
acts as an aggregation pheromone, which has been reported in (b)
other Platypus (Renwick et al., 1977, Shore snd Mclean, 1983) factor df MS F-ratio P
rather than as a sex pheromone (Audino, et al., 2005), such as lure 2 1.878 33.161 < 0.0001
test site 2 0.048 0.848 0.452
sulcatol and sulcatone for P. mutatus. Results of the field trap
test period 3 0.172 3.039 0.071
test in 2004 showed that racemic quercivorol I traps attracted lure × test site 4 0.105 1.860 0.182
more beetles than ethanol traps or control traps, as shown in test site × test period 6 0.073 1.285 0.334
the three-way ANOVAs and the post-hoc test (Table 1, 2). The test period × lure 6 0.085 1.503 0.258
effects of lure and test sites were highly significant. In addition, 12 0.057
test periods and lure × site interactions were nearly significant
using all test site data (Table 1a). Using the three sites where we Teble 2. F-ratio (df =1, 22) (above diagonal) and P-value (below
diagonal) for each pairwise comparison between lure type
were able to collect enough numbers of beetles, only the effect with the Bonferroni adjusting method. Number of beetles
of lure was significant and all interactions between factors were at two sites at Yamagata and one site at Kyoto were used.
not significant (Table 1b). We conducted a post-hoc test for each quercivorol ethanol water
pair of lures with the Bonferroni adjusting method, and found a quercivorol* - 19.337 49.917
significant difference between racemic quercivorol I and other ethanol 0.0003 - 3.469
lures (Table 2). control <0.0001 0.1582 -
*racemic quercivorol I
On the results of field trap test at Atsumi Yamagata in

Bulletin of FFPRI, Vol.6, No.1, 2007

54 TOKORO M.et al.

During 2006 of test period, we had much rain in spite of

field trpping at the test sites in Yamagata, the captured numbers
of the beetles by racemic quercivorol II in 2006 was much
less than that by racemic quercivorol I in 2004. Because of
their low density, we were not able to capture sufficient beetles
for statistical analyses. However, (1S,4R)-isomer showed the
most luring ability in the field. It showed concordance with
electrophysiological activity to (1S,4R)-isomer of GC–EAD
analysis, and we suggest that the (1S,4R)-isomer, quercivorol is
the main component of the aggregation pheromone. Quercivorol
is confirmed as novel pheromone component in insects (Witzgall
et al., 2004). Quercivorol was found in several kinds of plant
Fig. 4. Chemical structure of aggregation pheromone of P. quersivorus. leaves as an essential oil (Tabanca, et al., 2001; Ciccio et al.,
(1S,4R)-p-menth-2-en-1-ol (quercivorol) 2002; Albuquerque, et al., 2004; Sadyrbekov et al., 2006). To
develop methods for mass trapping (Lei & Bakke, 1981) and
communication disruption (Payne, 1981) against damage to
oak forests by P. quercivorus, we need further analyses for the

14 14
Number of beetles captured
Number of beetles captured

Yamagata (Site1) Yamagata (Site1)

12 Yamagata (Site2)
12 Yamagata (Site2)
Kyoto (Site1)
Kyoto (Site1)
Kyoto (Site2) Kyoto (Site2)

10 10

8 Female 8
Male

6 6

4 4

2 2
0 0 0 0 0 0 0
0 0
Quercivoro�� Ethanol Control Quercivorol � Ethanol Control

Fig. 5. Results of field trap tests at four sies in Yamagata (Kushibiki) and Kyoto (Keihoku) in 2004. Number
of females (mean ± SE) and males captured by traps with lures per 4 days during test period of 28
days. For details, see text and tables 1 and 2.

6
40
Number of beetles captured/ week
Number of beetles captured / week

1st week
1st week 2nd week

35 2nd week
5

30 4
25
3
20
2
15
1
10
0 0 0
5 0
(1S,4R) (1R,4S) Racemic Control
0 0
0 20 60 200 600 Fig. 7. Results of field trap tests at Yamagata (Oguni) in 2006.
Number of beetles captured by traps with lures per
Dose of racemic quercivorol � (�l / trap)
week during two weeks of test period.
(1S,4R) : 20 μl of (1S,4R)-isomer (quercivorol)
Fig. 6. Results of field trap tests at Yamagata (Atsumi) in 2005. (1R,4S) : 20 μl of (1R,4S)-isomer
Number of female and male beetls captured by traps Racermic: racemic quercivorol II (79.8 %, contains 4.1
with different doses per week during two weeks. % of trans-isomer)
For details, see text. Control : 50 ml of distilled water.

森林総合研究所研究報告 第 6 巻 1 号 , 2007
 Novel aggregation pheromone, (1S,4R)-p-menth-2-en-1-ol, of the ambrosia beetle, Platypus quercivorus (Coleoptera: Platypodidae)
sub-component of the pheromone and combined effect of the
pheromone and host kiromone component.
Acknowledgment
We appreciate the help of Ms. Ai Nozaki (Kyoto
Prefectural Forestry Experimental Station) who supplied
insects and collected boring frass. We thank Mr. Toshio Arihara
(Fukushima Forest Research Center) for his assistance with
collecting logs. Sincere gratitude is also extended to Drs.
Kiyoshi Nakamuta and Richard P. Shefferson (Forestry and
Forest Products Research Institute), for their critical reading of
the manuscript and valuable advice. This work was supported
in part by a Grant-in-Aid “Research Project for Utilizing
Advanced Technologies in Agriculture, Forestry and Fisheries”
from the Research Council, Ministry of Agriculture, Forestry
and Fisheries of Japan, grant No.1775.
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森林総合研究所研究報告 第 6 巻 1 号 , 2007
 Novel aggregation pheromone, quercivorol from the boring frass of an ambrosia beetle, Platypus quercivorus (Coleoptera: Platypodidae) 57

養菌性キクイムシ、カシノナガキクイムシ（鞘翅目：ナガキクイムシ科）
の集合フェロモン、(1S,4R) -p- メント -2- エン -1- オール

所 雅彦１)*、小林 正秀２)、斉藤 正一３)、衣浦 晴生４)、中島 忠一１)、

正田・加賀谷 悦子１)、柏木 丈拡５)、手林 慎一５)、金 哲史５)、森 謙治６）

要旨
 カシノナガキクイムシの集合フェロモンを、未交尾雄が穿入口付近に付着させた初期フラスの揮発性成
分から、ガスクロマトグラフ触角電図法により分析した。ガスクロマトグラフ質量分析計による分析等の
結果、ガスクロマトグラフ触角電図法により活性のあった成分の化学構造は（1S,4R)-p- メント -2- エン
-1- オール（ケルキボロール）と決定された。ケルキボロールとその光学異性体を用いた野外試験の結果、
この成分が集合フェロモンであることが証明された。

キーワード：カシノナガキクイムシ、集合フェロモン、 (1S,4R)-p- メント -2- エン - 1 - オール、

ケルキボロール、養菌性キクイムシ

＊ 森 林 総 合 研 究 所 森 林 昆 虫 研 究 領 域   〒 305-8687 茨 城 県 つ く ば 市 松 の 里 １ e-mail:tokoro@affrc.go.jp
1)森林総合研究所 森 林 昆 虫 研 究 領 域 (FFPRI)
2)京都府林業試験場
3)山形県森林研究研修センター
4)森 林 総 合 研 究 所 関 西 支 所 (FFPRI)
5)高知大学農学部
6)東洋合成工業 ( 株 ) 感光材研究所

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